Using single-species and algal communities to determine long-term adverse effects of silver nanoparticles on freshwater phytoplankton.

The physical and chemical properties of silver nanoparticles (AgNPs) have led to their increasing use in various fields such as medicine, food, and industry. Evidence has proven that AgNPs cause adverse effects in aquatic ecosystems, especially when the release of Ag is prolonged in time. Several studies have shown short-term adverse effects of AgNPs on freshwater phytoplankton, but few studies have analysed the impact of long-term exposures on these populations. Our studies were carried out to assess the effects of AgNPs on growth rate, photosynthesis activity, and reactive oxygen species (ROS) generation on the freshwater green algae Scenedesmus armatus and the cyanobacteria Microcystis aeruginosa, and additionally on microcystin (MC-LR) generation from these cyanobacteria. The tests were conducted both in single-species cultures and in phytoplanktonic communities exposed to 1 ngL-1 AgNPs for 28 days. The results showed that cell growth rate of both single-species cultures decreased significantly at the beginning and progressively reached control-like values at 28 days post-exposure. This effect was similar for the community-cultured cyanobacteria, but not for the green algae, which maintained a sustained decrease in growth rate. While gross photosynthesis (Pg) increased in both strains exposed in single cultures, dark respiration (R) and net photosynthesis (Pn) decreased in S. armatus and M. aeruginosa, respectively. These effects were mitigated when both strains were exposed under community culture conditions. Similarly, the ROS generation shown by both strains exposed in single-species cultures was mitigated when exposure occurred in community cultures. MC-LR production and release were significantly decreased in both single-species and community exposures. These results can supply helpful information to further investigate the potential risks of AgNPs and ultimately help policymakers make better-informed decisions about their utilization for environmental restoration.

Adverse effects of iron-based nanoparticles on freshwater phytoplankton Scenedesmus armatus and Microcystis aeruginosa strains.

Zero-valent nano-iron particles (nZVI) are increasingly present in freshwater aquatic environments due to their numerous applications in environmental remediation. However, despite the broad benefits associated with the use and development of nZVI nanoparticles, the potential risks of introducing them into the aquatic environment need to be considered. Special attention should be focused on primary producer organisms, the basal trophic level, whose impact affects the rest of the food web. Although there are numerous acute studies on the acute effects of these nanoparticles on photosynthetic primary producers, few studies focus on long-term exposures. The present study aimed at assessing the effects of nZVI on growth rate, photosynthesis activity, and reactive oxygen activity (ROS) on the freshwater green algae Scenedesmus armatus and the cyanobacteria Microcystis aeruginosa. Moreover, microcystin production was also evaluated. These parameters were assessed on both organisms singly exposed to 72 h-effective nZVI concentration for 10% maximal response for 28 days. The results showed that the cell growth rate of S. armatus was initially significantly altered and progressively reached control-like values at 28 days post-exposure, while M. aeruginosa did not show any significant difference concerning control values at any time. In both strains dark respiration (R) increased, unlike net photosynthesis (Pn), while gross photosynthesis (Pg) only slightly increased at 7 days of exposure and then became equal to control values at 28 days of exposure. The nZVI nanoparticles generated ROS progressively during the 28 days of exposure in both strains, although their formation was significantly higher on green algae than on cyanobacteria. These data can provide additional information to further investigate the potential risks of nZVI and ultimately help decision-makers make better informed decisions regarding the use of nZVI for environmental remediation.

Interference of heavy metals on the photosynthetic response from a Cr(VI)-resistant Dictyosphaerium chlorelloides strain.

The successful selection of a particular type of bioelement and its association to the appropriate transducer determines the specificity of a biosensor. Therefore, from a strain of chloroficea Dictyosphaerium chlorelloides, modified in laboratory to tolerate high Cr(VI) concentrations, the possible interferences of other heavy metals on photosynthetic activity were studied. After exposing wild type and Cr(VI)-resistant cells to increasing Ag(+1), Co(+2), Hg(+2), Cr(+3), Cu(+2), Zn(+2), Fe(+3) and Cd(+2) concentrations, both photosynthetic quantum yields was compared. Photosynthetic electron transport rates were measured with a TOXY-PAM chlorophyll fluorometer, non-linear regression analysis of each of the toxicity tests was done, and means of both groups were compared using unpaired t test. The results show no significant differences between both cell types when they were exposed to Ag(+1), Co(+2), Hg(+2), Cr(+3), Cu(+2), Fe(+3) and Cd(+2) metal ions, and extremely significant differences (p < 0.0001) to Zn(+2) exposures. These results demonstrate the suitability of this Cr(VI)-resistant type D. chlorelloides strain as a suitable bioelement to be coupled to a biosensor based on dual-head microalgae strategy to detect and quantify Cr(VI) in water courses and waste water treatment plants. However, some disturbance may be expected, especially when certain analyte species such as zinc are present in water samples tested. The analysis of binary mixtures between Zn(+2) and other heavy metals showed a slight antagonistic phenomenon in all cases, which should not alter the potential Zn(+2) interference in the Cr(+6) detection process.

Toxic risk associated with sporadic occurrences of Microcystis aeruginosa blooms from tidal rivers in marine and estuarine ecosystems and its impact on Artemia franciscana nauplii populations.

Microcystis aeruginosa is a species of freshwater cyanobacteria which can form harmful algal blooms in freshwater water bodies worldwide. However, in spite its sporadic occurrences for short periods of time in estuarine waters, their influence on zooplankton populations present in these ecosystems has not been extensively studied. In this work, Artemia franciscana was used as test organism model, studying mortality against several strains of M. aeruginosa with different degrees of toxigenicity, measuring whole-live cells and homogenate extracts. Results were compared with microcystin-LR equivalent content, measured by immunoassay. The results show that there were no significant differences between both exposure models (whole cells and extracts), and there are significant differences respect to the toxigenicity of cyanobacterial blooms depending of the M. aerugionosa strain involved in the process. Analysis of microcystin-LR equivalent concentration test immediately below the lowest significant concentration in all M. aerugionosa strains was used to determine the potential risk associated with the cell densities during a bloom. Comparison among the selected M. aerugionsa strains show that these factors have influence in the results obtained, and thus, several differences have been evidenced depending of the microcystin-LR equivalent production and the strain type involved.

Importance of strain type to predict the toxicological risk associated with Microcystis aeruginosa blooms: comparison of Microtox(®) analysis and immunoassay.

The occurrence of toxic cyanobacterial blooms in aquatic environments, associated with human health problems and animal deaths, has increased the need for rapid, reliable and sensitive methods to determine the toxicity of microcystin produced by cyanobacteria. An in vitro Microtox(®) system and a commercially available microcystin ELISA were used to screen out the potential risk associated with selected Microcystis aeruginosa strains (Ma1D-Ma8D). Results showed the existence of three differentiated groups in the selected M. aeruginosa strains. Strains Ma7D and Ma6D were determined to be very toxic, strains Ma2D, Ma1D and Ma5D as moderately toxic and strains Ma8D, Ma4D and MA3D as non-toxic. These results agreed with the microcystin concentration values obtained by immunoassay. Although the data obtained by other authors clearly show that Microtox(®) is not sensitive to microcystins, our results suggested that this bioluminescence assay may prove useful in the preliminary screening of cyanobacterial blooms for microcystin-based toxicity. Additionally, the combination of immunodetection and toxicity-based Microtox(®) provides a useful addition to the methods already available for detection of cyanobacterial toxins.

Toxic effects and specific chromium acquired resistance in selected strains of Dyctiosphaerium chlorelloides.

Due to its various uses, chromium contamination has become widespread in a diverse array of environments. The present study was carried out to investigate the toxic effect of chromium exposures on sensitive and resistant strains of the green algae Dyctiosphaerium chlorelloides, and to determine the nature and mechanism of chromium-resistant cells that arise. The toxic effect on the photosynthetic performance of chromium exposures in both cell populations, and the sensitive differences due to chromium oxidation state, were estimated, and the results indicate that although the photosynthetic performance in both strains were inhibited, there are not significant differences among IC(50(72)) values obtained in toxicity assays with both chromium oxidation states in wild-type cells, and however these differences are very significant when the assays were performed with Cr(VI) resistant cells. The 72-h 50% inhibitory concentration values obtained with Cr(III) exposures were similar for both strains. Additionally, by means of the SEM/EDX and TEM microscopic techniques, the occurrence of rapid morphological evolution in the microalgal cells and the possible detoxificant mechanisms was observed after exposure of the wild strain to chromium hexavalent. Moreover, the different response in photosynthetic activity observed between sensitive and resistant cells of D. chlorelloides in the presence of Cr(VI) and Cr(III) could be used to obtain a chromium-specific eukaryotic microalgal biosensor.

Toxic effects induced by salt stress on selected freshwater prokaryotic and eukaryotic microalgal species.

In order to determine the short-term impact induced by salt stress, cultures of Dictyosphaerium chlorelloides and Microcystis aeruginosa were grown in presence of increasing sea-salt concentrations. Growth rate and photosystem II activity in D. chlorelloides, and photosynthetic oxygen production (in both species) were analyzed. A concentration-dependent response was obtained with the presence of sea-salt in culture medium, being M. aeruginosa (EC(50(72)) = 76.6 mM) more sensitive to salt stress than D. chlorelloides (EC(50(72)) = 340.7 mM). However, comparative analysis between growth and Phi(PSII) inhibition in D. chlorelloides shown that there are not significant differences among EC(50(72)) values obtained. An immediate toxic response, induced by increase of sea-salt concentration, has been obtained applying the calculated EC(50(72)) values in both species. These results shown that sea-salt acts as a sensitive and rapid toxic compound in algal cells, and that the sensitivity of M. aeruginosa to salinity stress is much higher than that of D. chlorelloides.

Inhibition of growth and photosynthesis of selected green microalgae as tools to evaluate toxicity of dodecylethyldimethyl-ammonium bromide.

The effect of dodecylethyldimethyl-ammonium bromide (DEAB), a quaternary ammonium, compound widely used as disinfectant, on phytoplankton of inland water systems was analysed by using an experimental model. A toxicity test was based on inhibition of photosynthesis performances (effective quantum yield from photosystem II, Phi(PSII) and O(2) production) of the phytoplanktonic species Scenedesmus intermedius and Dictiosphaerium chlorelloides (Chlorophyceae) under growing doses of DEAB. A concentration-dependent toxic response was obtained in both parameters analysed. In addition, this response was almost immediate. Consequently, the measurement of both parameters (Phi(PSII )and O(2) production) allows to assess DEAB toxicity with higher standards of precision and repeatability. We propose that this procedure could be used to detect presence of quaternary ammonium pollutants in freshwater.